Project Summary/Abstract: Flow cytometry, by enabling high-throughput single-cell analysis of cell populations and particulates, has become essential instrumentation for many areas of NIH-funded biomedical research. The technology uses laser excitation of fluorescent tags to measure levels of labeled targets on cells, vesicles, or other particles in suspension. In parallel with traditional flow cytometry, the development of the CyTOF mass cytometer has also allowed powerful single cell analyses. Mass cytometers enable measurement of more than 40+ unique parameters per cell using rare metal labels identifiable by their unique atomic masses. However, mass cytometry is both far slower than traditional cytometry, and precludes use of cells in downstream applications. Despite this, mass cytometry publications have highlighted the advantages and insights possible with high-parameter analyses. This has incentivized continued development of traditional flow cytometry to enable higher dimensional measurements. This Shared Instrumentation Grant application is to request funding for purchase of a new flow cytometer analyzer to be placed in the Shared FACS Facility (a shared resource lab) at Stanford University. Since facility first offered researchers access to flow cytometer instrumentation more than 20 years ago, interest in pushing the limits of multi-parameter assays has been constant. In the intervening years since the facility?s last purchase of an analyzer almost 10 years ago, more than 15 new polymer-based dyes have been commercially released. These new fluorochromes include the Brilliant Violet?, Brilliant UltraViolet?, and Brilliant Blue? dyes. Current analyzers within the shared facility are unable to utilize the full array of reagents now available, and in addition, are severely over-scheduled, limiting researcher access. The Becton Dickinson FACSymphony is a state-of-the-art analyzer with 5-laser excitation lines capable of 30-parameter analysis. Purchase of this instrument will allow researchers to utilize a full-spectrum of commercially available dyes to identify phenotypic and functional properties of samples. With this state-of-the-art flow cytometers, more than 15 Major Users will have access to an instrument that will enhance their ability to answer complex questions heart disease, cancer, infection, allergy, and arthritis. With 30 available parameters, this high-throughput analyzer will contribute significantly to investigations into a variety of biomedical projects to further our understanding of healthy and disease states.